ChIP-Atlas: SRX5253409

Antigen

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Lysates were clarified from sonicated nuclei and protein-DNA complexes were isolated with anti-FOXB1 antibody. Approximately 10 – 15 ng CHIP DNA (around 15 μL) for each sample was used for CHIP-seq library construction following manufacturer instructions for NEBNext® UltraTM II DNA Libarry Prep Kit for Illumina (New England Biolabs). In brief, CHIP DNA was ended repaired at 20oC for 30 min followed by incubation at 65oC for 30 min. Ligation of adaptor (NEBNext Adaptor for Illumina) was then performed at 20oC for 15 min followed by U excision at 37oC for 15 min. Adaptor ligated and U-excised DNA was then cleaned up using AMPure XP beads (Beckman Coulter), washed twice with freshly prepared 80% ethanol and eluted in 17 μL of 10 mM Tris-HCl. PCR amplification was used to add the following indices GTCCGC, GTGAAA, GTGGCC, GTTTCG, CGTACG, and GAGTGG (NEBNExt Index 18 to 23) to the cleaned up DNA followed by another clean up step with AMPure XP Beads as described above. DNA was eluted in 30 μL 0.1X TE. The amount and quality of DNA was measured on Qubit 3.0 Fluorometer (Life Technologies) using Qubit™ dsDNA HS Assay Kit and Bioanalyzer (Agilent) using High Sensititivity DNA chip.